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Seeds of wheat (Triticum aestivum L. cv. Olgeurumil) were infected with Penicillium sp. at mean infection rate of 83%. Penicillium sp. was detected in endosperm with bran but not in embryo. Gaseous chlorine dioxide (ClO2) effectively inhibited growth of Penicillium sp. at concentration of 5 to 20 mg/ml. As treatment duration was extended from 1 to 3 h, growth of Penicillium sp. was completely suppressed even at 10 mg/ml. There was no significant reduction in the incidence of Penicillium sp. at 30% relative humidity (RH). However, the incidence of Penicillium sp. was 27.7% at 50% RH, further those were 3.5% and 0.2% at 70% and 80% RH, respectively. Seed germination was not affected by ClO2 treatment at all the RH conditions. Water-soaked seeds (30% seed moisture content) showed a drastic reduction in the incidence of Penicillium sp. when treated at more than 10 mg/ml of ClO2. The incidences of Penicillium sp. were 3.3, 1.8 and 1.2% at 10, 15 and 20 mg/ml, respectively. The incidence of Penicillium sp. in dry seeds with 9.7% seed moisture content did not reduce when treated with 5 and 10 mg/ml at 50% RH although it tended to decrease as ClO2 concentration increased to 20 mg/ml. Seed germination was not affected by ClO2 treatment at the tested concentrations. These results indicated that gaseous ClO2 was effective disinfectant to wheat seeds infected with Penicillium sp. and that the effectiveness of ClO2 strongly increased when moisture content around or inside of the seed was increased.